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ZFIN ID: ZDB-FIG-150811-31
Vanhollebeke et al., 2015 - Tip cell-specific requirement for an atypical Gpr124- and Reck-dependent Wnt/β-catenin pathway during brain angiogenesis. eLIFE   4 Full text @ Elife
ADDITIONAL FIGURES
EXPRESSION / LABELING:
Genes:
Fish:
Conditions:
Knockdown Reagents:
Anatomical Terms:
Stage Range: Prim-5 to Protruding-mouth
PHENOTYPE:
Fish:
Conditions:
Knockdown Reagents:
Observed In:
Stage Range: Prim-5 to Protruding-mouth

Fig. 3 Wnt/β-catenin signaling is controlled by Gpr124 and Reck.

(A) Maximal intensity projection of a confocal z-stack of Tg(7xTCF-Xia.Siam:GFP) Wnt/β-catenin reporter expression during brain vascular development in wild-type, gpr124 or reck morphant Tg(kdrl:ras-mCherry) embryos. (B) Activity of the endothelial-specific Wnt/β-catenin reporter in the PHBC and BA in wild-type, gpr124 or reck morphant Tg(fli1:Myr-mcherry) embryos at 40 hpf. (C) Lateral view of Tg(7xTCF-Xia.Siam:GFP) Wnt/β-catenin reporter expression at 54 hpf in the trunk region of wild-type, gpr124 or reck morphant Tg(sox10:mRPF) embryos (anterior to the left). (D) Quantification of hindbrain CtAs at 60 hpf after genetic and pharmacological inhibition of Wnt/β-catenin signaling. Heat-shock and pharmacological inhibition were performed as described in ‘Methods’. (E) Quantification of ngn1:GFP+ DRGs at 72 hpf after genetic inhibition of Wnt/β-catenin signaling. ngn1:GFP+ DRGs were counted on one side of the larvae. (F) Maximal intensity projection of a confocal z-stack of the cranial vasculature and quantification of hindbrain CtAs in control or gpr124 morphant Tg(kdrl:EGFP) embryos after exposure to the indicated GSK-3β inhibitors from the 16-somite stage onwards. Red arrows point to CtAs. Pharmacological inhibitions were performed as described in ‘Methods’. (G) Dorsal views and quantification of DRGs in control or gpr124 morphant Tg(ngn1:GFP);Tg(sox10:mRPF) embryos after exposure to the indicated GSK-3β inhibitors from the 16-somite stage onwards. ngn1:GFP+ DRGs were counted on one side of the larvae. Pharmacological inhibitions were performed as described in ‘Methods’. Scale bars, 50 µm. In all panels, values represent means ± SD (*p < 0.05; **p < 0.01; Kruskal–Wallis test: (D), (F), and (G); Mann–Whitney test: (E)).

Gene Expression Details
Gene Antibody Fish Conditions Stage Qualifier Anatomy Assay
EGFP ncv15Tg; ncv1Tg; nkuasgfp1aTg control Prim-25 basilar artery endothelial cell IFL
Prim-25 primordial hindbrain channel endothelial cell IFL
ncv15Tg; ncv1Tg; nkuasgfp1aTg + MO1-adgra2 standard conditions Prim-25 Not Detected basilar artery endothelial cell IFL
Prim-25 Not Detected primordial hindbrain channel endothelial cell IFL
ncv15Tg; ncv1Tg; nkuasgfp1aTg + MO1-reck standard conditions Prim-25 Not Detected basilar artery endothelial cell IFL
Prim-25 Not Detected primordial hindbrain channel endothelial cell IFL
s843Tg control Long-pec brain vasculature IFL
Long-pec central artery IFL
Long-pec cranial vasculature IFL
s843Tg + MO1-adgra2 control Long-pec Not Detected central artery IFL
Long-pec cranial vasculature IFL
s843Tg + MO1-adgra2 chemical treatment: 1-azakenpaullone Long-pec brain vasculature IFL
Long-pec central artery IFL
Long-pec cranial vasculature IFL
s843Tg + MO1-adgra2 chemical treatment: lithium chloride Long-pec brain vasculature IFL
Long-pec central artery IFL
Long-pec cranial vasculature IFL
GFP ia4Tg; s896Tg control Prim-5 brain IFL
Prim-5 primordial hindbrain channel endothelial cell IFL
Prim-15 brain IFL
Prim-15 primordial hindbrain channel endothelial cell IFL
Long-pec brain IFL
Long-pec central artery endothelial cell IFL
Long-pec primordial hindbrain channel endothelial cell IFL
ia4Tg; s896Tg + MO1-adgra2 standard conditions Prim-5 brain IFL
Prim-5 Not Detected primordial hindbrain channel endothelial cell IFL
Prim-15 brain IFL
Prim-15 Not Detected primordial hindbrain channel endothelial cell IFL
Long-pec brain IFL
Long-pec Not Detected central artery endothelial cell IFL
Long-pec Not Detected primordial hindbrain channel endothelial cell IFL
ia4Tg; s896Tg + MO1-reck standard conditions Prim-5 brain IFL
Prim-5 Not Detected primordial hindbrain channel endothelial cell IFL
Prim-15 brain IFL
Prim-15 Not Detected primordial hindbrain channel endothelial cell IFL
Long-pec brain IFL
Long-pec Not Detected central artery endothelial cell IFL
Long-pec Not Detected primordial hindbrain channel endothelial cell IFL
ia4Tg; vu234Tg control Long-pec dorsal root ganglion IFL
ia4Tg; vu234Tg + MO1-adgra2 standard conditions Long-pec Not Detected dorsal root ganglion IFL
ia4Tg; vu234Tg + MO1-reck standard conditions Long-pec Not Detected dorsal root ganglion IFL
sb1Tg; vu234Tg control Protruding-mouth dorsal root ganglion IFL
Protruding-mouth neuron IFL
sb1Tg; vu234Tg + MO1-adgra2 control Protruding-mouth Not Detected dorsal root ganglion IFL
Protruding-mouth neuron IFL
sb1Tg; vu234Tg + MO1-adgra2 chemical treatment: 1-azakenpaullone Protruding-mouth dorsal root ganglion IFL
Protruding-mouth neuron IFL
sb1Tg; vu234Tg + MO1-adgra2 chemical treatment: lithium chloride Protruding-mouth dorsal root ganglion IFL
Protruding-mouth neuron IFL
mCherry ia4Tg; s896Tg control Prim-5 primordial hindbrain channel endothelial cell IFL
Prim-15 basilar artery endothelial cell IFL
Prim-15 primordial hindbrain channel endothelial cell IFL
Long-pec central artery endothelial cell IFL
Long-pec primordial hindbrain channel endothelial cell IFL
ia4Tg; s896Tg + MO1-adgra2 standard conditions Prim-5 primordial hindbrain channel endothelial cell IFL
Prim-15 primordial hindbrain channel endothelial cell IFL
Long-pec Not Detected central artery endothelial cell IFL
Long-pec primordial hindbrain channel endothelial cell IFL
ia4Tg; s896Tg + MO1-reck standard conditions Prim-5 primordial hindbrain channel endothelial cell IFL
Prim-15 primordial hindbrain channel endothelial cell IFL
Long-pec Not Detected central artery endothelial cell IFL
Long-pec primordial hindbrain channel endothelial cell IFL
ncv15Tg; ncv1Tg; nkuasgfp1aTg control Prim-25 basilar artery endothelial cell IFL
Prim-25 primordial hindbrain channel endothelial cell IFL
ncv15Tg; ncv1Tg; nkuasgfp1aTg + MO1-adgra2 standard conditions Prim-25 basilar artery endothelial cell IFL
Prim-25 primordial hindbrain channel endothelial cell IFL
ncv15Tg; ncv1Tg; nkuasgfp1aTg + MO1-reck standard conditions Prim-25 basilar artery endothelial cell IFL
Prim-25 primordial hindbrain channel endothelial cell IFL
mRFP ia4Tg; vu234Tg control Long-pec dorsal root ganglion IFL
ia4Tg; vu234Tg + MO1-adgra2 standard conditions Long-pec dorsal root ganglion IFL
ia4Tg; vu234Tg + MO1-reck standard conditions Long-pec dorsal root ganglion IFL
sb1Tg; vu234Tg control Protruding-mouth dorsal root ganglion IFL
Protruding-mouth glial cell IFL
sb1Tg; vu234Tg + MO1-adgra2 control Protruding-mouth Not Detected dorsal root ganglion IFL
Protruding-mouth glial cell IFL
sb1Tg; vu234Tg + MO1-adgra2 chemical treatment: 1-azakenpaullone Protruding-mouth dorsal root ganglion IFL
Protruding-mouth glial cell IFL
sb1Tg; vu234Tg + MO1-adgra2 chemical treatment: lithium chloride Protruding-mouth dorsal root ganglion IFL
Protruding-mouth glial cell IFL
Antibody Labeling Details No data available
Phenotype Details
Fish Conditions Stage Phenotype
ia4Tg; s896Tg + MO1-adgra2 standard conditions Prim-5 primordial hindbrain channel Wnt signaling pathway decreased process quality, abnormal
Prim-15 primordial hindbrain channel Wnt signaling pathway decreased process quality, abnormal
Long-pec hindbrain central artery absent, abnormal
Long-pec hindbrain central artery decreased amount, abnormal
Long-pec primordial hindbrain channel Wnt signaling pathway decreased process quality, abnormal
ia4Tg; s896Tg + MO1-reck standard conditions Prim-5 primordial hindbrain channel Wnt signaling pathway decreased process quality, abnormal
Prim-15 primordial hindbrain channel Wnt signaling pathway decreased process quality, abnormal
Long-pec hindbrain central artery absent, abnormal
Long-pec hindbrain central artery decreased amount, abnormal
Long-pec primordial hindbrain channel Wnt signaling pathway decreased process quality, abnormal
ia4Tg; vu234Tg + MO1-adgra2 standard conditions Long-pec dorsal root ganglion Wnt signaling pathway decreased process quality, abnormal
ia4Tg; vu234Tg + MO1-reck standard conditions Long-pec dorsal root ganglion Wnt signaling pathway decreased process quality, abnormal
ncv15Tg; ncv1Tg; nkuasgfp1aTg + MO1-adgra2 standard conditions Prim-25 basilar artery Wnt signaling pathway decreased process quality, abnormal
Prim-25 primordial hindbrain channel Wnt signaling pathway decreased process quality, abnormal
ncv15Tg; ncv1Tg; nkuasgfp1aTg + MO1-reck standard conditions Prim-25 basilar artery Wnt signaling pathway decreased process quality, abnormal
Prim-25 primordial hindbrain channel Wnt signaling pathway decreased process quality, abnormal
s843Tg chemical treatment: IWR-1-endo Pec-fin central artery decreased amount, abnormal
s843Tg + MO1-adgra2 control Long-pec central artery absent, abnormal
s843Tg; w32Tg heat shock Pec-fin central artery decreased amount, abnormal
s843Tg; w35Tg heat shock Pec-fin central artery decreased amount, abnormal
sb1Tg; vu234Tg + MO1-adgra2 control Protruding-mouth dorsal root ganglion decreased amount, abnormal
sb1Tg; vu234Tg + MO1-adgra2 chemical treatment: 1-azakenpaullone Protruding-mouth dorsal root ganglion amount, normal
sb1Tg; vu234Tg + MO1-adgra2 chemical treatment: lithium chloride Protruding-mouth dorsal root ganglion amount, normal
sb1Tg; w35Tg heat shock Protruding-mouth dorsal root ganglion decreased amount, abnormal
Acknowledgments:
ZFIN wishes to thank the journal eLIFE for permission to reproduce figures from this article. Please note that this material may be protected by copyright. Full text @ Elife